Optimization of Laboratory Diagnostics of Primary Biliary Cholangitis: When Solid-Phase Assays and Immunofluorescence Combine

Federica Gaiani; Roberta Minerba; Alessandra Picanza; Annalisa Russo; Alessandra Melegari; Elena De Santis; Tommaso Trenti; Lucia Belloni; Silvia Peveri; Rosalia Aloe; Carlo Ferrari; Luigi Laghi; Gian Luigi de'Angelis; Chiara Bonaguri

doi:10.3390/jcm11175238

Optimization of Laboratory Diagnostics of Primary Biliary Cholangitis: When Solid-Phase Assays and Immunofluorescence Combine

J Clin Med. 2022 Sep 5;11(17):5238. doi: 10.3390/jcm11175238.

Authors

Federica Gaiani^{1

2}, Roberta Minerba³, Alessandra Picanza³, Annalisa Russo³, Alessandra Melegari⁴, Elena De Santis⁴, Tommaso Trenti⁵, Lucia Belloni⁶, Silvia Peveri⁷, Rosalia Aloe³, Carlo Ferrari^{1

8}, Luigi Laghi^{1

2}, Gian Luigi de'Angelis^{1

2}, Chiara Bonaguri³

Affiliations

¹ Department of Medicine and Surgery, University of Parma, Via Gramsci 14, 43126 Parma, Italy.
² Gastroenterology and Endoscopy Unit, University Hospital of Parma, Via Gramsci 14, 43126 Parma, Italy.
³ Laboratory of Clinical Chemistry and Hematology, University Hospital of Parma, Via Gramsci 14, 43126 Parma, Italy.
⁴ Autoimmunity Unit, Department of Laboratory Medicine and Pathology, S. Agostino Estense Hospital, Via Giardini 1355, 41126 Baggiovara, Italy.
⁵ Department of Laboratory Medicine and Pathology, S. Agostino Estense Hospital, Via Giardini 1355, 41126 Baggiovara, Italy.
⁶ Unit of Clinical Immunology, Allergy and Advanced Biotechnologies, Azienda Unità Sanitaria Locale-IRCCS of Reggio-Emilia, Viale Risorgimento 80, 42123 Reggio-Emilia, Italy.
⁷ Allergology Unit, Guglielmo da Saliceto Hospital, Via Giuseppe Taverna 49, 29121 Piacenza, Italy.
⁸ Laboratory of Viral Immunopathology, Unit of Infectious Diseases and Hepatology, University Hospital of Parma, Via Gramsci 14, 43126 Parma, Italy.

Abstract

The laboratory diagnostics of primary biliary cholangitis (PBC) have substantially improved, thanks to innovative analytical opportunities, such as enzyme-linked immunosorbent assays (ELISA) and multiple immunodot liver profile tests, based on recombinant or purified antigens. This study aimed to identify the best diagnostic test combination to optimize PBC diagnosis. Between January 2014 and March 2017, 164 PBC patients were recruited at the hospitals of Parma, Modena, Reggio-Emilia, and Piacenza. Antinuclear antibodies (ANA) and anti-mitochondrial antibodies (AMA) were assayed by indirect immunofluorescence (IIF), ELISA, and immunodot assays (PBC Screen, MIT3, M2, gp210, and sp100). AMA-IIF resulted in 89.6% positive cases. Using multiple immunodot liver profiles, AMA-M2 sensitivity was 94.5%, while anti-gp210 and anti-sp100 antibodies were positive in 16.5% and 17.7% of patients, respectively. PBC screening yielded positive results in 94.5% of cases; MIT3, sp100, and gp210 were detected by individual ELISA test in 89.0%, 17.1%, and 18.9% of patients, respectively. The association of PBC screening with IIF-AMA improved the diagnostic sensitivity from 89.6% to 98.2% (p < 0.01). When multiple immunodot liver profile testing was integrated with AMA-IIF, the diagnostic sensitivity increased from 89.1% to 98.8% (p < 0.01). The combination of IIF with solid-phase methods significantly improved diagnostic efficacy in PBC patients.

Keywords: anti-mitochondrial antibodies; indirect immunofluorescence; primary biliary cholangitis; solid-phase assay.

Grants and funding

Funding was provided by the Programma di Ricerca Regione-Università 2010–2012-Area 1—Strategic Research Programmes Regione Emilia-Romagna Progetto Triennale, under the project name: “A Tailored Approach to Immune-Monitoring and Clinical Management of Viral and Auto-Immune Diseases.”